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Title: Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library

Abstract

Recombinant EstE1 protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was then crystallized at 290 K by the hanging-drop vapour-diffusion method. EstE1, a new thermostable esterase, was isolated by functional screening of a metagenomic DNA library from thermal environment samples. This enzyme showed activity towards short-chain acyl derivatives of length C4–C6 at a temperature of 303–363 K and displayed a high thermostability above 353 K. EstE1 has 64 and 57% amino-acid sequence similarity to est{sub pc}-encoded carboxylesterase from Pyrobaculum calidifontis and AFEST from Archaeoglobus fulgidus, respectively. The recombinant protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was crystallized at 290 K by the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 2.3 Å resolution from an EstE1 crystal; the crystal belongs to space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 73.71, c = 234.23 Å. Assuming the presence of four molecules in the asymmetric unit, the Matthews coefficient V{sub M} is calculated to be 2.2 Å{sup 3} Da{sup −1} and the solvent content is 44.1%.

Authors:
 [1];  [2];  [3];  [1];  [3];  [1];  [2]
+ Show Author Affiliations
  1. Department of Biology, Yonsei University, Seoul 120-749 (Korea, Republic of)
  2. (Korea, Republic of)
  3. Department of Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of)
Publication Date:
OSTI Identifier:
22356264
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 2; Other Information: PMCID: PMC2150951; PMID: 16511287; PUBLISHER-ID: fw5062; OAI: oai:pubmedcentral.nih.gov:2150951; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; AFFINITY; CRYSTALLIZATION; CRYSTALS; DIFFUSION; ENVIRONMENT; ESCHERICHIA COLI; HISTIDINE; IRON; MOLECULES; RESOLUTION; SCREENING; SPACE GROUPS; STRAINS; X-RAY DIFFRACTION

Citation Formats

Byun, Jung-Sue, Protein Network Research Center, Yonsei University, Seoul 120-749, Rhee, Jin-Kyu, Kim, Dong-Uk, Oh, Jong-Won, Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr, and Protein Network Research Center, Yonsei University, Seoul 120-749. Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106000832.
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Byun, Jung-Sue, Protein Network Research Center, Yonsei University, Seoul 120-749, Rhee, Jin-Kyu, Kim, Dong-Uk, Oh, Jong-Won, Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr, & Protein Network Research Center, Yonsei University, Seoul 120-749. Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library. United Kingdom. doi:10.1107/S1744309106000832.
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Byun, Jung-Sue, Protein Network Research Center, Yonsei University, Seoul 120-749, Rhee, Jin-Kyu, Kim, Dong-Uk, Oh, Jong-Won, Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr, and Protein Network Research Center, Yonsei University, Seoul 120-749. Wed . "Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library". United Kingdom. doi:10.1107/S1744309106000832.
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@article{osti_22356264,
title = {Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library},
author = {Byun, Jung-Sue and Protein Network Research Center, Yonsei University, Seoul 120-749 and Rhee, Jin-Kyu and Kim, Dong-Uk and Oh, Jong-Won and Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr and Protein Network Research Center, Yonsei University, Seoul 120-749},
abstractNote = {Recombinant EstE1 protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was then crystallized at 290 K by the hanging-drop vapour-diffusion method. EstE1, a new thermostable esterase, was isolated by functional screening of a metagenomic DNA library from thermal environment samples. This enzyme showed activity towards short-chain acyl derivatives of length C4–C6 at a temperature of 303–363 K and displayed a high thermostability above 353 K. EstE1 has 64 and 57% amino-acid sequence similarity to est{sub pc}-encoded carboxylesterase from Pyrobaculum calidifontis and AFEST from Archaeoglobus fulgidus, respectively. The recombinant protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was crystallized at 290 K by the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 2.3 Å resolution from an EstE1 crystal; the crystal belongs to space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 73.71, c = 234.23 Å. Assuming the presence of four molecules in the asymmetric unit, the Matthews coefficient V{sub M} is calculated to be 2.2 Å{sup 3} Da{sup −1} and the solvent content is 44.1%.},
doi = {10.1107/S1744309106000832},
journal = {Acta Crystallographica. Section F},
number = Pt 2,
volume = 62,
place = {United Kingdom},
year = {Wed Feb 01 00:00:00 EST 2006},
month = {Wed Feb 01 00:00:00 EST 2006}
}
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Journal Article:
DOI:  10.1107/S1744309106000832
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